GSM7049635: liver scRNA-seq; Mus musculus; RNA-Seq
Minced tissue was placed into 10 mL of an enzyme solution containing 1 mg/mL collagenase type IV and 10 U/mL of DNase with shaking for 1 hour at 37 °C. Tissue was filtered through 70 μm cell strainer, flushed with 10 mL PBS, and centrifuged for 5 minutes at 380 g. After elimination of red blood cells by hypotonic lysis, leukocytes were isolated by centrifugation at 890 g for 15 minutes at 20°C through 3 mL Lymphoprep and 4 mL of cell suspension in DPBS. Single cell libraries are generated using the 10X Genomics platform.  First, Feature Barcode conjugated molecules are bound to cell surface proteins. Cells are loaded into the Chromium Controller where they are partitioned into nanoliter-scale Gel Beads-in-emulsion with a single barcode per cell.  Reverse transcription is performed, and the resulting cDNA is amplified. The full-length amplified cDNA is used to generate libraries.